1.Expression of cystic fibrosis transmembrane conductance regulator in rat ovary.
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(5):584-7
The protein expression of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl(-) channel, in ovarian stimulated premature female rat ovary during a cycle of follicle development and corpus luteum formation was investigated. Animals were injected with 10 U pregnant Mare's serum gonadotropin (PMSG) and subsequently 10 U hCG 48 h later. Time-dependent immunohistochemistry and Western blotting experiments were performed before and 24, 48, 72 h after hCG treatment. The immunohistochemistry revealed that administration of PMSG stimulated the CFTR expression in thecal cell layer and granulosa cell layer of mature follicles 48 h post injection, coincident with the PMSG-induced peak in follicular estradiol. However, the expression of CFTR in the granulose lutein cell layer and thecal lutein cell layer was time-dependently reduced following hCG injection, in accordance with the gradually increased progestogen level during luteum corpus formation. Western blotting analysis demonstrated that rat ovarian tissue expressed the special CFTR band at 170 kD. It is concluded that cAMP-dependent Cl(-) channels are involved in regulation of follicle development and luteum formation.
Connective Tissue Growth Factor/genetics
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Connective Tissue Growth Factor/*metabolism
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Connective Tissue Growth Factor/*physiology
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Corpus Luteum/growth & development
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Ovarian Follicle/growth & development
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Ovary/*metabolism
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Rats, Wistar
3.Research advances on ADAM28 expression and ADAM28-mediated tumor metastasis.
Xiao-Lu ZHU ; Qian-Ming WANG ; Fei-Er FENG ; Xing-Long ZHENG ; Xiao-Hui ZHANG
Journal of Experimental Hematology 2014;22(4):1142-1147
A disintegrin-metalloproteinase 28 (ADAM28) is one of important members of ADAM family, that is involved in various biological events including cell adhesion, proteolysis, growth and metastasis of solid tumors and hematological malignancies. Studies have shown that ADAM28 is highly expressed in several human tumors, such as lung, breast and bladder cancers, and chronic lymphocytic leukemia, and its tissue expression levels correlate with cancer metastasis. ADAM28-mediated cancer cell metastasis may be related with the cleavage of von Willebrand's factor (vWF), insulin-like growth factor binding protein-3 (IGFBP-3) and connective tissue growth factor (CTGF), as well as the promoting PSGL-1/P-selectin-mediated cell adhesion. This review summarizes the basic and translational aspects of ADAM28 biology that might stimulate the interest in ADAM28 research and discovery of novel ADAM28 targets, providing potential novel therapies for metastatic cancers.
ADAM Proteins
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metabolism
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Cell Adhesion
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Connective Tissue Growth Factor
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metabolism
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Humans
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Insulin-Like Growth Factor Binding Protein 3
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metabolism
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Neoplasm Metastasis
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Neoplasms
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pathology
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von Willebrand Factor
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metabolism
4.Organic anion transporter system mediates the transport of aristolochic acid in tubular epithelial cells.
Yan-Yan WANG ; Yi-Pu CHEN ; Hong-Liang RUI
Acta Academiae Medicinae Sinicae 2009;31(4):476-480
OBJECTIVETo investigate whether aristolochic acid can be transported into human kidney proximal tubular cell (HKC) and its potential mechanism.
METHODSIntracellular aristolochic acid was measured by liquid chromatography-tandem mass spectrometry. The release of lactate dehydrogenase (LDH) induced by aristolochic acid in the presence of organic anion transporter inhibitor (probenecid) or organic cation transporter inhibitor (tetraethylammonium) was evaluated. The effects of probenecid on aristolochic acid induced connective tissue growth factor (CTGF) mRNA and protein expression were also examined by real time polymerase chain reaction and Western blot, respectively.
RESULTSAristolochic acid was detected in the suspension of the denatured HKC after incubation with aristolochic acid sodium salt. The release of LDH from HKC, which was induced by 60 mg/L aristolochic acid sodium salt, was significantly inhibited by 1 mmol/L probenecid (P < 0.01), but not by 1 mmol/L tetraethylammonium. The increased CTGF mRNA and protein expression in HKC stimulated by 40 mg/L aristolochic acid sodium salt was significantly down-regulated by 1 mmol/L probenecid (P < 0.05), with an inhibition rate of 16% and 21%, respectively.
CONCLUSIONAristolochic acid can be transported into HKC by organic anion transport system, and then exerts its biological effects.
Aristolochic Acids ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Epithelial Cells ; metabolism ; Humans ; Kidney ; physiology ; Organic Anion Transporters ; metabolism
6.Experimental study on expression of connective tissue growth factor in viral myocarditis in mice.
Jing-hui SUN ; Zhen ZHANG ; Shu-bo ZHAI ; Yu-tong ZHANG
Chinese Journal of Pediatrics 2011;49(10):782-787
OBJECTIVETo study the expression of connective tissue growth factor (CTGF) in the myocardial tissue of mice with viral myocarditis (VMC).
METHODBalb/c mice were infected with coxsackie virus B3 (CVB3) to establish VMC model. The mice were divided into control group (n = 50) and VMC group (n = 50). on days 4, 7, 14 and 21 after infection, heart specimens of 8 mice were randomly taken and examined after HE staining for myocardial necrosis and cellular infiltration. The area of positive Masson stained myocardium collagen fibers was measured, and collagen volume fraction (CVF) was measured. Then the level of serum creatine phosphokinase-MB (CKMB) was determined. The levels of CTGF and TGF-β₁ were detected by streptavidin peroxidase immunoperoxidase technique. Expression of CTGF and TGF-β₁ were detected with reverse transcription-polymerase chain reaction (RT-PCR). At the same time, the correlations were analyzed.
RESULT(1) The level of CKMB peaked on day 7, and decreased afterwards (455.45 ± 37.95, 606.95 ± 35.64, 573.62 ± 42.90, 308.60 ± 20.49, respectively, 4 - 21 d points), in which 4, 7, 14 d points, there was significant difference compared with control group (t = 6.144, 12.558, 11.182, respectively, P < 0.01). (2) CVF increased significantly on day 14 (8.22 ± 1.95, t = 4.486, P < 0.01) and day 21 (9.46 ± 1.87, t = 4.486, P < 0.01) in VMC group. (3) Measured by streptavidin peroxidase immunoperoxidase technique, the levels of CTGF (171.50 ± 10.25, 141.70 ± 10.863, 110.35 ± 11.051, 81.05 ± 10.190, respectively, 4 - 21 d points) and TGF-β₁ (184.90 ± 11.480, 150.25 ± 9.915, 103.50 ± 10.455, 84.15 ± 9.848, respectively, 4 - 21 d points) increased after day 4 in VMC (P < 0.01). (4) Measured by RT-PCR, the expression of CTGF mRNA and TGF-β₁ increased in VMC group, and the increase was enhanced with the disease development (P < 0.01). (5) The expression of CTGF and TGF-β₁ was positively linearly correlated (r = 0.987, P < 0.01), the expression of CTGF was negatively correlated with CVF (r = -0.901, P < 0.01), but the expression of CTGF was detected earlier than myocardial fibrosis.
CONCLUSIONThe increase of CTGF expression was associated with the severity of myocardial fibrosis in VMC. These results suggest that abnormal expression of CTGF may take part in the development of fibrosis in VMC.
Animals ; Connective Tissue Growth Factor ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; pathology ; virology ; Myocardium ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism
7.Changes of TGF-β1 and CTGF in rats with increased blood flow-induced pulmonary artery hypertension.
Rong ZHU ; Liang HE ; Junmei XU ; Yanling ZHANG ; Yongbin HU
Journal of Central South University(Medical Sciences) 2012;37(10):1013-1020
OBJECTIVE:
To evaluate the role of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) in the pathogenesis of pulmonary artery hypertension, we observed the dynamic expression of TGF-β1 and CTGF in rats with high blood flow.
METHODS:
Fifty adult male SD rats were randomly divided into 5 groups: a sham group (group S) and groups with right pneumonectomy for 1, 2, 4 and 6 weeks (PE1, PE2, PE4 and PE6 group), 10 rats per group. The mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were measured. TGF-β1 and CTGF protein expression in the lung tissues were determined with immunohistochemistry and Western blot. The expression of TGF-β1 mRNA and CTGF mRNA in the lung tissues was evaluated by RT-PCR.
RESULTS:
Compared with group S, the mPAP and RVHI in the rats were significantly increased in group PE1, PE2, PE4, and PE6 (P<0.05); the indicators of vascular remodeling [(MA+PMA)%, RMT, and RMA] were markedly elevated in group PE4 and PE6 (P<0.05), but not in group PE1 and PE2. Immunohistochemical staining of TGF-β1 and CTGF were more prominent in all of the right pneumonectomy groups than in the sham group. Western blot showed that the level of TGF-β1 protein was significantly increased in all of the right pneumonectomy groups (P<0.01), and the peak was observed in group PE2, whereas the level of CTGF protein was markedly elevated in group PE4 and PE6 (P<0.05), but no change was noticed in group PE1 and PE2. Compare to group S, the mRNA level of TGF-β1 was dramatically increased in all right pneumonectomy groups (P<0.01), peaked at group PE2, and remained high in group PE4 and PE6. In contrast, the elevation of mRNA level of CTGF was not significant in group PE1, but group PE2, PE4 and PE6 demonstrated significant mRNA level of CTGF (P<0.01). Correlation analysis showed that the protein and mRNA levels of CTGF were positively correlated with RMT and RMA ( r=0.743, r=0.906; P<0.05), while no correlation between the protein and mRNA level of TGF-β1 with RMT or RMA. There was no correlation between the mRNA level of TGF-β1 and CTGF.
CONCLUSION
TGF-β1 and CTGF play a role in the pathogenesis of increased pulmonary blood flow-induced pulmonary hypertension.
Animals
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Connective Tissue Growth Factor
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metabolism
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Hypertension, Pulmonary
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metabolism
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Lung
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metabolism
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Male
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Pulmonary Artery
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RNA, Messenger
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Rats
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Rats, Sprague-Dawley
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Transforming Growth Factor beta1
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metabolism
8.Expression of connective tissue growth factor and its down-regulation by simvastatin administration in pulmonary hypertensive rats.
Bin LIU ; Xian-min WANG ; Tong-fu ZHOU ; Yi-min HUA ; Han-min LIU ; Li WEI ; Li-na QIAO ; Xiao-qin WANG ; Shan-shan ZHAO ; Kun SHI
Chinese Journal of Pediatrics 2008;46(5):359-365
OBJECTIVETo explore the role of expression of connective tissue growth factor (CTGF) in pulmonary vascular remodeling of pulmonary hypertensive rats, and investigate the regulation of CTGF expression by simvastatin in this animal model.
METHODSEighty male Sprague-Dawley rats (350 to 400 g) were randomized to 7 groups. The rats in group PM(1 - 21) (n = 10) and PM(1 - 35) (n = 12) were treated with pneumonectomy + monocrotaline (MCT), and sacrificed at the 21st or 35th experimental day;those in groups PMS(1 - 35) (n = 12), PMS(21 - 35) (n = 12), PMV(1 - 35) (n = 12) and PMV(21 - 35) (n = 12) were given daily lavage of simvastatin (or vehicle) as intervention measure which began from the 1st and 21st experimental days, respectively; additional 10 rats were used as control without any intervention. The animals were sacrificed at the end of experiment (35 th day) as hemodynamic measurements and study on the morphological parameters relevant to pulmonary vascular remodeling were performed on each group of rats. The expression of ET-1 mRNA, CTGF mRNA and protein, and synthesis of collagen in these pneumonectomized, MCT-treated rats were compared between control and rats treated with simvastatin.
RESULTSRats in PM(1 - 35) Group developed severe PAH (mPAP = 39.75 +/- 3.62 mm Hg) (1 mm Hg = 0.133 kPa), right ventricular hypertrophy [RV/(LV + S) ratio = 0.627 +/- 0.040], and arterial medial hypertrophy (WT% = 61.73 +/- 5.39), these parameters of the control animals were 17.10 +/- 1.20 mm Hg, 0.262 +/- 0.018 and 14.71 +/- 1.16, respectively. CTGF mRNA and protein were mainly located in pulmonary arterial smooth muscle cells and interstitial macrophage shown by in situ hybridization and immunohistochemistry, respectively. The expression of ET-1 mRNA and CTGF mRNA detected by fluorescent quantitative RT-PCR in Group PM(1 - 35) were significantly increased in comparison with controls, and so did the CTGF protein expression determined by Western blotting in these diseased rats. The content of hydroxyproline (1.30 +/- 0.19 microg/mg wet lung) was remarkably higher than that of control animals (0.56 +/- 0.10 microg/mg wet lung). The up-regulation of ET-1 and CTGF gene expression, and elevated synthesis of hydroxyproline were reversed in rats intervened with simvastatin. The pulmonary hypertension, right ventricular hypertrophy and medial hypertrophy were attenuated in all simvastatin-treated rats no matter the intervention was initiated from the beginning or midway of the study.
CONCLUSIONThe up-regulation of CTGF gene expression may play an important role in the development of pulmonary vascular remodeling in PAH. Simvastatin can prevent and, to some extent, reverse the vascular remodeling via down-regulation of CTGF gene expression.
Animals ; Connective Tissue Growth Factor ; metabolism ; Down-Regulation ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Simvastatin ; pharmacology
9.A study on the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles.
Huiling LI ; Jinwen CHEN ; Xuan XU ; Ruhao YANG ; Xudong XIANG ; Dongshan ZHANG
Journal of Central South University(Medical Sciences) 2016;41(2):113-120
OBJECTIVE:
To study the safety and efficiency of the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles, and to evaluate its potential clinical application.
METHODS:
The potential and conditions regarding the transfection self-made lipid microbubbles (CY5)-labeled-oligonucleotide (ODN) or CY5-labeled-ODN connective tissue growth factor (CTGF) into the rat kidney were evaluated. Th e safety was evaluated by HE staining, liver and renal function tests. The transfection efficiency was evaluated by fluorescence microscopy. Th e expression of CTGF was detected by RT-PCR and Western blot.
RESULTS:
Self-made lipid microbubble and/or ultrasound significantly enhanced the efficiency of gene transfer and expression in the kidney. Especially, 85%-90% of total glomerular could be transfected. CY5-labeled-ODN expression could be observed in glomerular, tubular and interstitial area. Th ere was no significant change in blood tests aft er gene transfer. Levels of LDH in 7 days were decreased compared with that at the fi rst day aft er the transfection (P<0.05). CTGF expression was successfully suppressed by transfection of CTGF-antisense-ODN into kidney.
CONCLUSION
The ultrasound-mediated gene transfer by self-made lipid microbubble could enhance the efficiency of ODN and expression in the rat kidney. Th is self-made lipid microbubbles supplement may be use for transfection of target genes.
Animals
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Connective Tissue Growth Factor
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genetics
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metabolism
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Kidney
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metabolism
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Lipids
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chemistry
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Microbubbles
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Oligonucleotides, Antisense
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genetics
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RNA, Messenger
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Rats
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Transfection
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Ultrasonics
10.Expressions of ATF3 and CTGF and their regulation by estradiol in the prepuce of hypospadias patients.
Juan ZHOU ; Hao WANG ; Yu-Bing PENG ; Qi CHEN ; Jun DA ; Zhong WANG ; Li-Min MA
National Journal of Andrology 2009;15(12):1075-1080
OBJECTIVEEstrogen is closely associated with hypospadias. The present study was to explore the molecular mechanism of hypospadias caused by estradiol.
METHODSFibroblasts obtained from the prepuce of hypospadiac and normal children were cultured in vitro and treated with 17-beta ethinyl estradiol (17-EE) at the concentrations of 1 micromol/L to 0.1 nmol/L for 2 hours, or at 0.1 micromol/L for 0.5, 1, 2, 4, 8, 16 and 24 hours. MTT assay was used to evaluate the effect of 17-EE on the proliferation of the cells, and RT-PCR was employed to detect the expressions of the activating transcription factor-3 (ATF3) and connective tissue growth factor (CTGF) in the hypospadiac tissue. The results were compared with those obtained from the nonhypospadiac tissue.
RESULTSThe expressions of ATF3 and CTGF were significantly upregulated in the hypospadiac tissue as compared with the nonhypospadiac group. At the concentration of 1 micromol/L, 17-EE significantly inhibited the proliferation of the cells. ATF3 mRNA was elevated at 1-2 hours, while CTGF mRNA showed no significant changes in 24 hours.
CONCLUSIONATF3 and CTGF are two candidate genes involved in the etiology of hypospadias. And estradiol may induce hypospadias by upregulating the expressions of ATF3 and CTGF.
Activating Transcription Factor 3 ; genetics ; metabolism ; Cells, Cultured ; Child ; Connective Tissue Growth Factor ; genetics ; metabolism ; Estradiol ; pharmacology ; Estrogens ; pharmacology ; Fibroblasts ; metabolism ; Foreskin ; metabolism ; Humans ; Hypospadias ; genetics ; metabolism ; Male